OBJECTIVE: To explore the peripheral neural mechanism underlying representation along the distribution of stomach meridian induced by intestinal inflammatory reaction using diarrhea predominant-irritable bowel syndrome (IBS-D) mice. METHODS: Among 62 healthy male C57BL/6 mice of clean grade, 12 mice were randomly selected and divided into a control group and a model group, 6 mice in each group, additionally, 12 mice were randomly selected and divided into a Tianshu group, a Liangqiu group and a Zusanli group, 4 mice in each group. In the model group, citrobacter was administered orally to establish IBS-D model. In the control group and the model group, the visceral pain threshold was observed using fecal colorectal distension (fCRD) induced electromyography of external oblique muscle, the positive cell number of neutrophil in the colonic muscularis was detected by myeloperoxidase (MPO) staining, the number, location and distribution rule of Evans blue (EB) extravasation points were observed by injection of EB staining solution into the tail vein. In the Tianshu group, the Liangqiu group and the Zusanli group, fluorescent dye Dil was injected at bilateral "Tianshu" (ST25), "Liangqiu" (ST34) and "Zusanli" (ST36) respectively, to observe the dye-positive cell number in different dorsal root ganglion (DRG) segments. In the control group and the model group, the activation of satellite glial cells (SGCs) in different DRG segments was observed by immunofluorescence. RESULTS: Compared with the control group, in the model group, the area under curve of electromyography of external oblique muscle was increased at fCRD of 25, 50 and 75 μL distilled water (P<0.001, P<0.01); the MPO-positive cell number of neutrophil in the colonic muscularis was increased (P<0.01). Few EB extravasation points could be found in the control group, while there were much more EB extravasation points observed in the model group, which was specially distribution in the area of stomach meridian, from "Huaroumen" (ST24) to "Zusanli" (ST36), as well as the surface area dominated by L₂-L₅ segment of the spinal cord. The Dil-positive cells were mainly exhibited in the DRG of T₁₁, L₅ and L₄ segments in the Tianshu group, the Liangqiu group and the Zusanli group, respectively. Compared with the control group, the ratio of glial fibrillary acidic protein (GFAP)/glutamine synthetase (GS) co-expression was increased in the DRG of T₁₁, L₄ and L₅ segments in the model group (P<0.05, P<0.01). CONCLUSION The activation of SGCs within DRG of T₁₁, L₄ and L₅ segments may relate closely to the occurrence of the representation along the stomach meridian distribution in IBS-D mice.
Animals ; Male ; Mice ; Irritable Bowel Syndrome/therapy* ; Mice, Inbred C57BL ; Meridians ; Stomach/physiopathology* ; Humans ; Acupuncture Points ; Disease Models, Animal

ObjectiveBased on ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， network pharmacology and molecular docking techniques， to explore the pharmacodynamic material basis and mechanism of Renshen Wuweizi Tang in treating spleen-lung Qi deficiency syndrome. MethodsThe chemical components in the decoction of Renshen Wuweizi Tang were systematically characterized and identified by UPLC-Q-TOF-MS/MS， and network pharmacology was used to screen potential active ingredients， collect component targets and gene sets related to spleen-lung Qi deficiency syndrome， and obtain protein interaction relationships through STRING. Cytoscape 3.9.1 was used to construct a "formula-syndrome" association network and calculate topological feature values. Gene ontology（GO） function and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis were performed on core genes to explore potential pharmacodynamic links， the average shortest path between the formula-drug target network and the pharmacodynamic link gene network was calculated to discover dominant pharmacodynamic links， and MCODE plugin was used to identify core gene clusters from the dominant pharmacodynamic links， which were validated using Gene Expression Omnibus（GEO）， and molecular docking was performed between key components and core targets. ResultsOne hundred and thirty-seven components were identified in the negative ion mode， and eighty components were identified in the positive ion mode. After deduplication， a total of 185 components were identified， mainly composed of triterpenoid saponins（49） and flavonoids（54）. Based on the "formula-syndrome" correlation network analysis， energy metabolism was determined to be the dominant pharmacodynamic link of Renshen Wuweizi Tang in the treatment of spleen-lung Qi deficiency syndrome. The results of molecular docking showed that 7 components（adenosine， atractylenolide Ⅱ， atractylenolide Ⅲ， ginsenoside Rg₁， glycyrrhizin B₂， glycyrrhizin E₂ and campesterol） from 4 medicinal materials（Ginseng Radix et Rhizoma， Atractylodis Macrocephalae Rhizoma， Glycyrrhizae Radix et Rhizoma and Poria） in this formula might regulate energy metabolism by acting on 6 targets， namely cyclic adenosine monophosphate-response element binding protein 1（CREB1）， glyceraldehyde-3-phosphate dehydrogenase（GAPDH）， interleukin（IL）-6， nuclear transcription factor（NF）-κB1， peroxisome proliferator-activated receptor α（PPARα）， and tumor necrosis factor（TNF）， thus improving the symptoms of diseases related to spleen-lung Qi deficiency syndrome. ConclusionThis study established a UPLC-Q-TOF-MS/MS for rapid characterization and identification of chemical components in the decoction of Renshen Wuweizi Tang， expanding the understanding of the material composition of this formula， and found that 7 components might act on the key advantageous pharmacodynamic link "energy metabolism" through 6 targets to improve the related symptoms of spleen-lung Qi deficiency syndrome. This can provide a reference for the subsequent exploration of the material benchmark and mechanism of the famous classical formula.

Objective:To establish a genomic nucleic acid mass spectrometry detection platform for allelic risk associated with Alzheimer's disease.Methods:Whole blood samples of 61 patients diagnosed as Alzheimer's disease in the Affiliated Brain Hospital of Guangzhou Medical University from December 28th, 2023 to 31st, March 2024 were collected and deoxynucleic acid (DNA) was extracted, including 22 males and 39 females, aged (67.36 ± 8.18) years old. After screening out 17 risk gene loci in Chinese population, multiplex polymerase chain reaction primers, single-base extension primers and Sanger sequencing primers were designed. Ten samples were used for primer optimization and debugging through Sanger sequencing and time-of-flight mass spectrometry to establish a detection system. The remaining samples were genotyped using a time-of-flight mass spectrometer and verified by Sanger sequencing for accuracy evaluation. Five samples were selected for gradient dilution and then subjected to time-of-flight mass spectrometry detection to evaluate the detection limit. Three clinical samples, one case of Escherichia coli and one case of Staphylococcus aureus genomic DNA samples were selected for cross-reaction research. The anti-interference ability of the detection system was evaluated against hemolysis, chylous substances and conventional anticoagulants in the samples. Two samples, one wild and one homozygous mutation sample with representative peak shapes, were selected to evaluate the anti-interference ability. Four samples containing the common genotypes of all gene loci in the system were selected and repeated 10 times to evaluate the precision.Results:The minimum intensity of single-base extension primers on mass spectrometry is greater than half of the maximum intensity. All 17 risk gene loci screened were successfully typed. The time-of-flight mass spectrometry detection results of 1,037 loci from 61 samples showed that the genotyping detection rate was 100%. The genotypes of the 20 DNA samples were completely consistent with the results of Sanger sequencing, with an accuracy rate of 100%. The mass spectrometry detection results of five samples after gradient dilution indicated that the low detection limit was 5 ng of DNA. The reaction system has a strong anti-interference ability against hemolysis of samples, chylous substances, conventional anticoagulants and DNA cross-contamination. Homologous allele interference and no cross-reaction between the bacterial genome and 17 gene loci do not affect the risk genome detection results. The results of 10 repeated mass spectrometry tests on 4 samples showed that the precision was 100%.Conclusion:The genomic detection system of Alzheimer's disease risk has been successfully established to provide an auxiliary mean for disease diagnosis and risk assessment.

Objective To establish a risk prediction scheme for recurrence of diabetic foot ulcer based on biochemical test indexes.Methods The clinical data of totally 198 patients with diabetic foot ulcer in the hos-pital from January 2016 to December 2019 were retrospectively analyzed,and randomly divided into a training group(n=139)and a verification group(n=59)according to a ratio of 7∶3.The general data of the training group and the verification group were compared.The training group was divided into recurrence group(n=75)and non-recurrence group(n=64)according to whether there was recurrence or not during following-up.General data of the recurrence group and the non-recurrence group were compared,and the influencing factors of recurrence of diabetic foot ulcer were analyzed by Logistic regression to establish a Nomogram model.Re-ceiver operating characteristic(ROC)curve was used to evaluate the prediction ability of the Nomogram mod-el for both the training group and the verification group,and the Nomogram model was internally verified by Bootstrap method.The clinical net benefit rate of this Nomogram model was verified by decision curve analy-sis(DCA).Results The recurrence rate of 198 patients was 53.03％(105/198).Glycosylated hemoglobin A1c(HbA1c,OR=1.86 6,95％CI:1.377-2.529),white blood cell count(WBC,OR=1.687,95％CI:1.094-2.602),C-reactive protein(CRP,OR=1.704,95％CI:1.145-2.537),platelet(PLT,OR=1.939,95％CI:1.270-2.961),serum creatinine(Scr,OR=1.687,95％CI:1.193-2.387),blood urea nitrogen(BUN,OR=1.685,95％CI:1.227-2.315),urine microalbumin-creatinine ratio(ACR,OR=1.842,95％CI:1.230-2.759),and vascular endothelial growth factor(VEGF,OR=1.829,95％CI:1.281-2.614)were risk factors for recurrence of diabetic foot ulcer(P＜0.05),albumin(ALB,OR=0.462,95％CI:0.287-0.742)and total bilirubin(TBIL,OR=0.506,95％CI:0.327-0.783)were protective factors for the recurrence of diabetic foot ulcer(P＜0.05).The area under the curve(AUG)of the Nomogram model was 0.928(95％CI:0.802-0.985),the sensitivity was 91.71％,and the specificity was 82.41％.The AUC of the prediction group was 0.775(95％CI:0.617-0.890),the sensitivity was 79.17％,and the specificity was 86.48％.Hosmer-Lemeshow goodness of fit test showed P values were both over 0.05 in the training group and the verification group,and the model calibration was well.The Nomogram model predicts that the training group could obtain net clinical benefits in the range of 0.00％to 96.00％,and the verification group can obtain net clinical bene-fits in the range of 0.00％to 95.00％.Conclusion HbA1c,WBC,CRP,PLT,Scr,BUN,ACR and VEGF are risk factors for the recurrence of diabetic foot ulcer,and ALB and TBIL are protective factors for the recur-rence of diabetic foot ulcer.Integrating these factors to construct a Nomogram model could predict the recur-rence of diabetic foot ulcer effectively.

Hashimoto's thyroiditis(HT)is an autoimmune thyroid disease characterized by diffuse enlargement of thyroid gland and elevated thyroid autoantibodies.HT is closely related to emotional factors,conveyance and dispersion function of the liver,and the qi movement of the five zang organs.This paper explores the approach to the differentiation and treatment of HT based on Professor Yang Shuyu's view of regulation of both mind and body.In the view of regulation of both mind and body,the regulation of mind is to harmonize emotions,and the regulation of body refers to the conveyance and dispersion function of zang-fu organs.Based on the view of regulation of both mind and body,the pathological changes of HT are characterized by the disordered conveyance and dispersion of qi movement,and the stable phase,hyperthyroid phase,and hypothyroid phase of HT correspond to obstructed conveyance and dispersion,excessive conveyance and dispersion,and insufficient conveyance and dispersion,respectively.The emotional disorders caused by the failure of mind regulation exist throughout the disease course.Therefore,the treatment of HT can be conducted by using the therapies of unblocking,suppressing,and tonifying separately for stable phase,hyperthyroid phase,and hypothyroid phase to address the root cause,and then the conveyance and dispersion of visceral qi movement are restored.Besides,the therapeutic method of resolution is used to alleviate symptoms by removing goiter and dissipating nodules.Simultaneously,emotional regulation therapy is incorporated to achieve a comprehensive efficacy for harmonizing physique-spirit and mind-body through the regulation of emotions,conveyance and dispersion of qi movement,and visceral functions.The view of regulation of both mind and body provides new methods and approaches for traditional Chinese medicine management of HT.

In 2022， Hainan provincial government launched the project for the prevention and control of viral hepatitis with the goals of a hepatitis B screening rate of 90%， a diagnostic rate of 90%， and a treatment rate of 80% among people aged 18 years and above by the year 2025， and the main intervention measures include population-based prevention， case screening， antiviral therapy， and health management. As of December 31， 2024， a total of 6.875 million individuals in the general population had been screened for hepatitis B， with a screening rate of 95.6%. A total of 184 710 individuals with positive HBsAg were identified， among whom 156 772 were diagnosed through serological reexamination， resulting in a diagnostic rate of 84.9%. A total of 50 742 patients with chronic hepatitis B were identified， among whom 42 921 had hepatitis B-specific health records established for health management， with a file establishment rate of 84.6%. A total of 31 553 individuals received antiviral therapy， with a treatment rate of 62.2%. A total of 2.503 million individuals at a high risk of hepatitis C were screened， among whom 4 870 tested positive for HCV antibody and 3 858 underwent HCV RNA testing， resulting in a diagnostic rate of 79.2%， and 1 824 individuals with positive HCV RNA were identified， among whom 1 194 received antiviral therapy， with a treatment rate of 65.5%. In addition， 159 301 individuals with negative HBsAg and anti-HBs and an age of 20 — 40 years were inoculated with hepatitis B vaccine free of charge. Through the implementation of the project for the prevention and control of viral hepatitis， a large number of hepatitis patients have been identified， treated， and managed in the province within a short period of time， which significantly accelerates the efforts to eliminate the crisis of viral hepatitis.

Objective To analyze the evaluation value of dynamic monitoring of changes in serum brain natriuretic peptide(BNP),urinary kidney injury molecule-1(KIM-1),and serum cystatin C(CysC)levels in acute kidney injury(AKI)after pediatric heart surgery.Methods 138 children who underwent cardiopulmonary bypass for congenital heart disease correction in our hospital from January 2023 to June 2024 were selected.The incidence of postoperative AKI in children was counted and they were divided into the AKI group and the non-AKI group accordingly.The changing levels of serum BNP,KIM-1,and CysC in the two groups of children were dynamically monitored and compared.The receiver operating characteristic curve(ROC)was used to evaluate the occurrence and severity of AKI after pediatric heart surgery.Results Among the 138 children,81 did not develop AKI and 57 developed AKI,with an incidence rate of 41.30%.In the AKI group,there were a mild group(n=20)and a moderate-severe group(n=37).Regarding the AKI group and the non-AKI group:there were differences in the levels of BNP,KIM-1,and CysC at 6 h,12 h,24 h,and 48 h after surgery(P<0.05).The levels of BNP,KIM-1,and CysC in the AKI group were higher than those in the non-AKI group(P<0.05),and there were differences in the changing trends of BNP,KIM-1,and CysC levels between the two groups(P<0.05).The results of multivariate Logistic regression analysis showed that BNP,KIM-1 and CysC were all factors influencing the occurrence of AKI after pediatric cardiac surgery(P<0.05).The ROC curve showed that the AUC of the three combined to predict the occurrence of AKI after pediatric heart surgery was 0.893,which was significantly higher than 0.723,0.812,and 0.761 of BNP,KIM-1,and CysC respectively.Regarding the mild group and the moderate-severe group:there were differences in the levels of BNP,KIM-1,and CysC at 6 h,12 h,24 h,and 48 h after surgery(P<0.05).The levels of BNP,KIM-1,and CysC in the mild group were lower than those in the moderate-severe group(P<0.05),and there were differences in the changing trends of BNP,KIM-1,and CysC levels between the two groups(P<0.05).The results of multivariate Logistic regression analysis indicated that BNP,KIM-1 and CysC were all factors influencing the severity of AKI after pediatric cardiac surgery(P<0.05).The ROC curve showed that the AUC of the three combined to evaluate the severity of AKI was 0.908,which was significantly higher than 0.780,0.762,and 0.720 of BNP,KIM-1,and CysC respectively.Conclusion Dynamic monitoring of changes in serum BNP,KIM-1,and CysC levels has a high evaluation value for the occurrence and severity of AKI after pediatric heart surgery,and the combined detection has a higher evaluation value.

Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylated β-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressing β-catenin ΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20 μmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10 μmol/L QCT was chosen as the intervention concentration.Overexpression of β-catenin ΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10 μmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10 μmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.

Objective:To evaluate the efficacy and safety of nebulized inhalation of recombinant human interferon (IFN) α1b injection in the treatment of respiratory syncytial virus (RSV) associated lower respiratory tract infections (pneumonia and bronchiolitis) in children.Methods:A randomized, double-blind, parallel, placebo-controlled add-on design was used.Children with pneumonia or bronchiolitis aged 2 months to 5 years who tested positive for RSV antigen within 72 hours of onset from 30 clinical trial sites including Beijing Children′s Hospital, Capital Medical University between February 2021 and December 2022 were included in this study and randomly divided into 2 groups at a ratio of 1∶1 based on a stratified-block method.Both groups received basic treatments such as cough control, asthma relieving, expectorant treatment, fever reduction, oxygen therapy, etc.The experimental group received additional nebulized inhalation of IFN α1b injection at a dose of 2.0 μg/(kg·time), twice a day.The control group received nebulized inhalation of placebo twice a day.Clinical efficacy was evaluated based on indicators such as the duration of clinical symptoms and signs, and the Kaplan-Meier method was used to calculate the median and 95% CI of the duration of clinical symptoms and signs.The Log-rank test was used to compared data between groups.Safety was assessed through the incidence of adverse reactions and laboratory tests, and the Chi-square test was used to analyze the difference between groups. Results:There were 123 children in the experimental group and 122 children in the control group.The median durations of all the 5 clinical symptoms and signs [including shortness of breath, wheezing, dyspnea (visible retractions), decreased transcutaneous oxygen saturation, and abnormal mental state] in the experimental group after treatment were slightly shortened than those in the control group [2.7 d(95% CI: 1.9-3.0 d)] vs.[2.9 d(95% CI: 2.6-3.6 d), P=0.027].The improvement in dyspnea (retractions) was especially pronounced in the experimental group, with a relief rate of 50.0% (0, 100%) on the first day of administration[compared with 0 (0, 50.0%) in the control group ( Z=2.002, P=0.025)].The median duration of dyspnea in the experimental group was nearly 1 day shorter than that in the control group [1.0 d(95% CI: 0.7-1.7 d) vs.1.8 d(95% CI: 1.0-2.5 d), P=0.046].There were no significant difference in hospital stay [6.0(5.0, 8.0) d vs.6.5(5.0, 8.0) d, Z=0.675, P=0.500], oxygen therapy duration [32.0(14.0, 96.3) h vs.39.0 (24.0, 83.2) h, Z=0.094, P=0.925], the recovery rate from clinical symptoms during treatment [(105/106, 99.1%) vs.(96/101, 95.0%)], and recurrence rate [(0/106, 0) vs.(2/101, 2.0%)] between the 2 groups (all P>0.05).However, the above-mentioned four indicators in the experimental group showed a trend of clinical benefits.The quantitative virus detection results showed that the RSV viral load in both groups decreased after treatment compared to before treatment.After 2 days of treatment, the decline rate of RSV viral load from the baseline was 0.90 lg copies/(mL·d) in the experimental group and 0.25 lg copies/(mL·d)in the control group, with a statistically significant difference ( P<0.05).Furthermore, there was no statistically significant difference in the incidence of adverse reactions between the 2 groups ( P>0.05).Importantly, no drug-related serious adverse reactions occurred in both groups. Conclusions:The nebulized inhalation therapy of IFN α1b demonstrates efficacy and safety in treating pediatric RSV associated lower respiratory tract infections.It particularly offers outstanding clinical therapeutic value for severe children.

Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylated β-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressing β-catenin ΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20 μmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10 μmol/L QCT was chosen as the intervention concentration.Overexpression of β-catenin ΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10 μmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10 μmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.